GFP-Tagged Human Gastric Carcinoma N87 Cells

Frozen Vials in a Dry Ice Package.

Frozen vial

General Information# Human Gastric Carcinoma N87 Cells (N87 cells) were established from the liver metastatic site of a male gastric carcinoma patient. GFP-Tagged Human Gastric Carcinoma N87 Cells (GFP-N87 cells) are selected from N87 cells after infected with lentiviruses expressing GFP with puromycin. The cells can be cultivated with Universal Full Growth Medium (cAP-01B) and they have a minimum population doubling capacity > 12 when cultured following the detailed protocol described below.
Characterization of the Cells#
1.: N87 cells are negative for HIV-1, HBV, HCV, and mycoplasma.
Product Usage# The cells are offered for Research Use Only.
Handling of Arriving Cells# When you receive the cells in a frozen vial, you can transfer the vial of cells into a -80AC freezer for short period storage or a liquid nitrogen tank for long term storage. Thaw the cells in a 37AC water bath, and then transfer the cells in a T25 flask pre-coated with human Fibronectin (20ug/ml, cAP-42) as described in details in Subculture Protocol.
Protocols for Thawing the cells and subculture#
A):Pre-coating of T25 flasks Add 1ml of Fibronectin (cAP-42) into one T25 flask and make sure whole surface of the flask is covered with the coating solution and leave flask at room temperature for overnight or 37Ë¿C for 1 hour. Dispose excessive fibronectin by aspiration. Rinsing the flask with PBS once before the flask is ready to be used.
B):Thaw the frozen cell vial in a 37C water bath first, and then transfer the cells into the pre-coated T25 flask with 10ml of cAP-01B medium, cells usually become confluent with 5-7 days.
C):To passage the cells, rinse the cells in a T25 flask with 5ml HBSS (RT) twice; then add 2ml Trypsin/EDTA (RT) (cAP-23) into one T25 flask; gently dispose the excessive Trypsin/EDTA solution within 20 seconds by aspiration.
D):Leave the T25 flask with the cells at RT or 37C for 1 min (most cells usually will detach from the surface within 1-2 mins; or monitor the cells under a microscope until most of cells become rounded up, and then gently tap the flask against the bench surface, and the cells will move on the surface of the flask when monitoring under microscope.
E):Add 5ml Trypsin Neutralization Buffer and spin down the cells with 800g centrifugation for 5 mins.
G):Re-suspend the cell pellet with 10 or 15ml Full medium and transfer 5 ml each into 2 or 3 pre-coated T25 flasks (for 1/2 to 1/3 subculture ratio).
H):Change medium every 2 or 3days and the cells usually become confluent within 7 days (when split at a 1/3 ratio).